2024-03-30T07:21:52Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/2082502020-05-29T11:59:43Zcom_10261_47com_10261_8col_10261_300
2020-04-20T09:14:33Z
urn:hdl:10261/208250
Fast Global Phosphoproteome Profiling of Jurkat T cells by HIFU-TiO2-SCX-LC-MS/MS
Carrera, Mónica
Cañas, Benito
López-Ferrer, Daniel
Phosphoproteomics
High-intensity focused ultrasound (HIFU)
TiO2
Strong cation exchange chromatography (SCX)
Proteomics
Mass spectrometry (MS)
Human Jurkat leukemia T cells
10 pages, 6 figures, 1 table
We propose a new workflow for fast phosphoproteome profiling. The workflow is based on the use of accelerated in-solution trypsin digestion under an ultrasonic field provided by high-intensity focused ultrasound (HIFU) combined with an inverse strategy based on TiO2 selective phosphopeptide enrichment, fractionation by strong cation exchange chromatography (SCX) and analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS) using a high-resolution mass spectrometer. The performance of the method was established for the global phosphoproteome analysis of un-stimulated human Jurkat leukemia T cells (E6.1). Using this accelerated workflow, 15,367 phosphorylation sites from 13,029 different phosphopeptides belonging to 3,163 different phosphoproteins were efficiently identified with high-throughput and reproducibility in less than 15 h. The functional analysis revealed significant phosphorylation-based networks that are implicated in immune function and tumor development pathways. The present strategy, HIFU-TiO2-SCX-LC-MS/MS, is the fastest analytical method reported to date for generating large-scale phosphoproteomics datasets (<15 h)
2020-04-20T09:14:33Z
2020-04-20T09:14:33Z
2017
artículo
Analytical Chemistry - Columbus 89(17): 8853-8862 (2017)
0003-2700
http://hdl.handle.net/10261/208250
10.1021/acs.analchem.7b01321
1520-6882
eng
Postprint
https://doi.org/10.1021/acs.analchem.7b01321
Sí
openAccess
American Chemical Society